Wang Weilan, Huang Chengmin, Luo Peng, Yao Jiang, Li Jie, Wang Wenxia, Liu Fengqin
Department of General Surgery, Changxing People's Hospital, The Second Affiliated Hospital of Zhejiang University School of Medical Changxing Campus, Changxing, Zhejiang, People's Republic of China.
Department of Medical Research, Shanghai Topgen Biomedical Technology Co., Ltd., Shanghai, People's Republic of China.
Onco Targets Ther. 2021 May 19;14:3281-3293. doi: 10.2147/OTT.S290270. eCollection 2021.
This study was to explore the biological roles and underlying mechanism of circRNA WD repeat domain 27 (circWDR27).
The expression of circWDR27, microRNA-215-5p (miR-215-5p) and tripartite motif containing 44 (TRIM44) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assays were employed to detect cell proliferation. Flow cytometry was used to determine cell apoptosis and cell cycle distribution. Cell migration and invasion abilities were examined by wound healing and transwell assays. The protein levels of matrix metalloproteinase 2 (MMP2), MMP9 and TRIM44 were analyzed by Western blot assay. The relationship between miR-215-5p and circWDR27 or TRIM44 was predicted by bioinformatics tools and confirmed using dual-luciferase reporter assay. Mouse xenograft model was established to examine the role of circWDR27 in vivo.
CircWDR27 and TRIM44 were highly expressed while miR-215-5p was lowly expressed in PTC tissues and cells. Knockdown of circWDR27 suppressed cell proliferation and metastasis and induced cell cycle arrest and apoptosis in PTC cells. Moreover, miR-215-5p was a direct target of circWDR27, and its inhibition reversed the suppressive effect of circWDR27 knockdown on PTC cell progression. In addition, miR-215-5p directly targeted TRIM44, and miR-215-5p exerted its anti-cancer role in PTC cells by targeting TRIM44. Furthermore, circWDR27 positively regulated TRIM44 expression by sponging miR-215-5p. Importantly, knockdown of circWDR27 suppressed tumor growth in vivo by upregulating miR-215-5p and downregulating TRIM44.
CircWDR27 accelerates PTC progression via regulating miR-215-5p/TRIM44 axis, providing a potential therapeutic target for PTC.
本研究旨在探讨环状RNA WD重复结构域27（circWDR27）的生物学作用及潜在机制。
采用定量实时聚合酶链反应（qRT-PCR）检测circWDR27、微小RNA-215-5p（miR-215-5p）和含三联基序蛋白44（TRIM44）的表达。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和集落形成试验检测细胞增殖。流式细胞术用于测定细胞凋亡和细胞周期分布。通过伤口愈合试验和Transwell试验检测细胞迁移和侵袭能力。采用蛋白质免疫印迹法分析基质金属蛋白酶2（MMP2）、MMP9和TRIM44的蛋白水平。利用生物信息学工具预测miR-215-5p与circWDR27或TRIM44之间的关系，并通过双荧光素酶报告基因试验进行验证。建立小鼠异种移植模型以检测circWDR27在体内的作用。
在甲状腺乳头状癌（PTC）组织和细胞中，circWDR27和TRIM44高表达，而miR-215-5p低表达。敲低circWDR27可抑制PTC细胞的增殖和转移，诱导细胞周期阻滞和凋亡。此外，miR-215-5p是circWDR27的直接靶点，其抑制作用可逆转敲低circWDR27对PTC细胞进展的抑制作用。此外，miR-215-5p直接靶向TRIM44，miR-215-5p通过靶向TRIM44在PTC细胞中发挥抗癌作用。此外，circWDR27通过吸附miR-215-5p正向调节TRIM44的表达。重要的是，敲低circWDR27可通过上调miR-215-5p和下调TRIM44抑制体内肿瘤生长。
circWDR27通过调节miR-215-5p/TRIM44轴促进PTC进展，为PTC提供了一个潜在的治疗靶点。
